CRF receptor antagonists and methods relating thereto

ABSTRACT

Compounds are disclosed which have utility in the treatment of a variety of disorders, including the treatment of disorders manifesting hypersecretion of CRF in a warm-blooded animals, including stroke. The compounds of this invention have the following structures:                    
     wherein n, m, R, R 1 , R 2 , X and Ar are as defined herein, including stereoisomes and pharmaceutically acceptable salts thereof.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.09/439,841 filed Nov. 12, 1999, now U.S. Pat. No. 6,348,466, which is acontinuation-in-part of U.S. patent application Ser. No. 09/400,744filed Sep. 21, 1999 (now abandoned), which is a continuation-in-part ofU.S. patent application Ser. No. 09/190,958 filed Nov. 12, 1998 (nowabandoned), which applications are incorporated herein by reference intheir entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates generally to CRF receptor antagonists, and tomethods of treating disorders by administration of such antagonists to awarm-blooded animal in need thereof.

2. Description of the Related Art

The first corticotropin-releasing factor (CRF) was isolated from ovinehypothalmi and identified as a 41-amino acid peptide (Vale et al.,Science 213:1394-1397, 1981). Subsequently, sequences of human and ratCRF were isolated and determined to be identical, but different fromovine CRF in 7 of the 41 amino acid residues (Rivier et al., Proc. Natl.Acad. Sci. USA 80:4851, 1983; Shibahara et al., EMBO J. 2:775, 1983).

CRF has been found to produce profound alterations in endocrine, nervousand immune system function. CRF is believed to be the majorphysiological regulator of the basal and stress-release ofadrenocorticotropic hormone (“ACTH”), β-endorphin, and otherpro-opiomelanocortin (“POMC”)-derived peptides from the anteriorpituitary (Vale et al., Science 213:1394-1397, 1981). Briefly, CRF isbelieved to initiate its biological effects by binding to a plasmamembrane receptor which has been found to be distributed throughout thebrain (DeSouza et al., Science 224:1449-1451, 1984), pituitary (DeSouzaet al., Methods Enzymol. 124:560, 1986; Wynn et al., Biochem. Biophys.Res. Comm. 110:602-608, 1983), adrenals (Udelsman et al., Nature319:147-150, 1986) and spleen (Webster, E. L., and E. B. DeSouza,Endocrinology 122:609-617, 1988). The CRF receptor is coupled to aGTP-binding protein (Perrin et al., Endocrinology 118:1171-1179, 1986)which mediates CRF-stimulated increase in intracellular production ofcAMP (Bilezikjian, L. M., and W. W. Vale, Endocrinology 113:657-662,1983). The receptor for CRF has now been cloned from rat (Perrin et al.,Endo 133(6):3058-3061, 1993), and human brain (Chen et al., PNAS90(19):8967-8971, 1993; Vita et al., FEBS 335(1):1-5, 1993). Thisreceptor is a 415 amino acid protein comprising seven membrane spanningdomains. A comparison of identity between rat and human sequences showsa high degree of homology (97%) at the amino acid level.

In addition to its role in stimulating the production of ACTH and POMC,CRF is also believed to coordinate many of the endocrine, autonomic, andbehavioral responses to stress, and may be involved in thepathophysiology of affective disorders. Moreover, CRF is believed to bea key intermediary in communication between the immune, central nervous,endocrine and cardiovascular systems (Crofford et al., J. Clin. Invest.90:2555-2564, 1992; Sapolsky et al., Science 238:522-524, 1987; Tilderset al., Regul. Peptides 5:77-84, 1982). Overall, CRF appears to be oneof the pivotal central nervous system neurotransmitters and plays acrucial role in integrating the body's overall response to stress.

Administration of CRF directly to the brain elicits behavioral,physiological, and endocrine responses identical to those observed foran animal exposed to a stressful environment. For example,intracerebroventricular injection of CRF results in behavioralactivation (Sutton et al., Nature 297:331, 1982), persistent activationof the electroencephalogram (Ehlers et al., Brain Res. 278:332, 1983),stimulation of the sympathoadrenomedullary pathway (Brown et al.,Endocrinology 110:928, 1982), an increase of heart rate and bloodpressure (Fisher et al., Endocrinology 110:2222, 1982), an increase inoxygen consumption (Brown et al., Life Sciences 30:207, 1982),alteration of gastrointestinal activity (Williams et al., Am. J.Physiol. 253:G582, 1987), suppression of food consumption (Levine etal., Neuropharmacology 22:337, 1983), modification of sexual behavior(Sirinathsinghji et al., Nature 305:232, 1983), and immune functioncompromise (Irwin et al., Am. J. Physiol. 255:R744, 1988). Furthermore,clinical data suggests that CRF may be hypersecreted in the brain indepression, anxiety-related disorders, and anorexia nervosa. (DeSouza,Ann. Reports in Med. Chem. 25:215-223, 1990). Accordingly, clinical datasuggests that CRF receptor antagonists may represent novelantidepressant and/or anxiolytic drugs that may be useful in thetreatment of the neuropsychiatric disorders manifesting hypersecretionof CRF.

The first CRF receptor antagonists were peptides (see, e.g., Rivier etal., U.S. Pat. No. 4,605,642; Rivier et al., Science 224:889, 1984).While these peptides established that CRF receptor antagonists canattenuate the pharmacological responses to CRF, peptide CRF receptorantagonists suffer from the usual drawbacks of peptide therapeuticsincluding lack of stability and limited oral activity. More recently,small molecule CRF receptor antagonists have been reported. For example,substituted 4-thio-5-oxo-3-pyyrazoline derivatives (Abreu et al., U.S.Pat. No. 5,063,245) and substituted 2-aminothiazole derivatives(Courtemanche et al., Australian Patent No. AU-A-41399/93) have beenreported as CRF receptor antagonists. These particular derivatives werefound to be effective in inhibiting the binding of CRF to its receptorin the 1-10 μM range and 0.1-10 μM range, respectively.

More recently, numerous small molecule CRR receptor antagonists havebeen proposed, including the compounds disclosed in the following patentdocuments: WO 94/13643, WO 94/13644, WO 94/13661, WO 94/13676, WO94/13677, WO 95/10506, WO 95/33750, WO 96/35689, WO 97/00868, WO97,35539, WO 97/35580, WO 97,35846, WO 97/44038, WO 98/03510, WO98/05661, WO 98/08846, WO 98/08847, WO 98/11075, WO 98/15543, WO98/21200 and WO 98/29413.

Due to the physiological significance of CRF, the development ofbiologically-active small molecules having significant CRF receptorbinding activity and which are capable of antagonizing the CRF receptorremains a desirable goal. Such CRF receptor antagonists would be usefulin the treatment of endocrine, psychiatric and neurologic conditions orillnesses, including stress-related disorders in general.

While significant strides have been made toward achieving CRF regulationthrough administration of CRF receptor antagonists, there remains a needin the art for effective small molecule CRF receptor antagonists. Thereis also a need for pharmaceutical compositions containing such CRFreceptor antagonists, as well as methods relating to the use thereof totreat, for example, stress-related disorders. The present inventionfulfills these needs, and provides other related advantages.

BRIEF SUMMARY OF THE INVENTION

In brief, this invention is generally directed to CRF receptorantagonists, and more specifically to CRF receptor antagonists havingthe following general structure (I):

including stereoisomers and pharmaceutically acceptable salts thereof,wherein m, n, X, R, R₁, R₂ and Ar are as defined below.

The CRF receptor antagonists of this invention have utility over a widerange of therapeutic applications, and may be used to treat a variety ofdisorders or illnesses, including stress-related disorders. Such methodsinclude administering an effective amount of a CRF receptor antagonistof this invention, preferably in the form of a pharmaceuticalcomposition, to an animal in need thereof. Accordingly, in anotherembodiment, pharmaceutical compositions are disclosed containing one ormore CRF receptor antagonists of this invention in combination with apharmaceutically acceptable carrier and/or diluent.

These and other aspects of the invention will be apparent upon referenceto the following detailed description. To this end, various referencesare set forth herein which describe in more detail certain procedures,compounds and/or compositions, and are hereby incorporated by referencein their entirety.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed generally to compounds useful ascorticotropin-releasing factor (CRF) receptor antagonists.

In a first embodiment, the CRF receptor antagonists of this inventionhave the following structure (I):

including stereoisomers and pharmaceutically acceptable salts thereof,

wherein:

n is 1 or 2;

m is 0, 1, 2 or 3;

X is N or CR′;

R is an optional substituent which, at each occurrence, is independentlyC₁₋₆alkyl, C₃₋₆alkenyl C₁₋₆ alkylidenyl or C₁₋₆alkylAr;

R′ is hydrogen, halogen or C₁₋₆alkyl;

R₁ is —C(H)_(0,1)(R₃)(R₄);

R₂ is hydrogen or C₁₋₆alkyl;

R₃ is hydrogen, keto, C₁₋₆alkyl, mono- or di(C₃₋₆cycloalkyl)methyl,C₃₋₆cycloalkyl, C₃₋₆alkenyl, hydroxyC₁₋₆alkyl,C₁₋₆alkylcarbonyloxyC₁₋₆alkyl, or C₁₋₆alkyloxyC₁₋₆alkyl, and

R₄ is hydrogen, Ar¹, C₁₋₆alkylAr¹, OAr¹, C₁₋₈alkyl, C₁₋₆alkyloxy,C₃₋₆cycloalkyl, mono- or di(C₃₋₆cycloalkyl)methyl, C₃₋₆alkenyl,C₃₋₆alkynyl, C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkoxyAr¹, hydroxyC₁₋₆alkyl,thienylC₁₋₆alkyl, furanylC₁₋₆alkyl, C₁₋₆alkylthioC₁₋₆alkyl, morpholinyl,mono- or di(C₁₋₆alkyl)aminoC₁₋₆alkyl, amino, (C₁₋₆alkyl)amino,di(C₁₋₆alkyl)amino, (C₁₋₆alkylAr¹)amino, (C₁₋₆alkyl)(Ar¹)amino,C₁₋₆alkylcarbonylC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,sulfonyl(C₁₋₈alkyl), C(═O)C₁₋₆alkyl, C₁₋₈alkyl substituted withphthalimide, Ar¹, OAr¹, NHAr¹, C(═O)Ar¹, C(═O)NHAr¹ or —C(═O)NH₂, or aradical of the formula —(C₁₋₆alkanediyl)-Y—(CO)_(0,1)—Ar¹ where Y is O,NH or a direct bond, or

R₃ and R₄ taken together with the carbon atom to which they are attachedform a C₅₋₈cycloalkyl, a C₅₋₈cycloalkenyl, a C₃₋₁₂heterocycle, phenyl,naphthyl, or a C₅₋₈cycloalkyl fused to Ar¹, each of which beingoptionally substituted with one or more substituents independentlyselected from C₁₋₆alkyl;

Ar is phenyl, naphthyl or an aromatic C₃₋₁₂heterocycle, each beingoptionally substituted with 1, 2 or 3 substituents independentlyselected from halo, C₁₋₆alkyl, trifluoromethyl, O(trifluoromethyl),hydroxy, cyano, C₁₋₆alkyloxy, phenoxy, benzoxy, C₁₋₆alkylthio, nitro,amino, mono- or di(C₁₋₆alkyl)amino, (C₁₋₆alkyl)(C₁₋₆alkanoyl)amino, orpiperidinyl, or wherein two substituents taken together are aC₁₋₆alkylidinyl or a C₁₋₆alkylidenyl having one, two or three carbonatoms replaced with a heteroatom individually selected from oxygen,nitrogen or and sulfur; and

Ar¹ is phenyl, naphthyl or an aromatic C₃₋₁₂heterocycle, each of whichbeing optionally substituted with 1, 2 or 3 substituents independentlyselected from halo, C₁₋₆alkyl, C₁₋₆alkyloxy, di(C₁₋₆alkyl)amino,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, trifluoromethyl sulfanyl(C₁₋₆alkyl), andC₁₋₆alkyl substituted with morpholinyl.

In the context of this invention, the preceding terms have the meaningsset forth below.

“Keto” represents ═O.

“C₁₋₆alkyl” or “C₁₋₈alkyl” represents a straight chain or branched alkylhaving from 1 to 6 carbon atoms or 1 to 8 carbon atoms, respectively,such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl,n-pentyl, and the like.

“C₁₋₆alkyloxy” represents the group —O(C₁₋₆alkyl), such as methoxy,ethoxy, and the like.

“C₁₋₆alkylthio” represents the group —S(C₁₋₆alkyl), such as —SCH₃,—SCH₂CH₃, and the like.

“C₃₋₆cycloalkyl” represents a cyclic alkyl having from 3 to 6 carbonatoms, including cyclopropyl, cyclopentyl, cyclopentyl, and cyclohexyl.

“C₅₋₈cycloalkyl” represents a cyclic alkyl having from 5 to 8 carbonatoms, such as cyclopentyl, cyclohexyl, and the like.

“C₅₋₈cycloalkenyl” represents a cyclic alkyl having from 5 to 8 carbonatoms an at least one double bond.

“C₃₋₆alkenyl” represents an unsaturated straight chain or branched alkylhaving from 3 to 6 carbon atoms, and having at least one double bond,such as propylenyl, 1-butenyl, 2-butenyl, 2-methylpropenyl, and thelike.

“C₃₋₆alkynyl” represents an unsaturated straight chain or branched alkylhaving from 3 to 6 carbon atoms, and having at least one triple bond,such as propylynyl, 1-butynyl, 2-butynyl, 2-methylpropynyl, and thelike.

“HydroxyC₁₋₆alkyl” represents a C₁₋₆alkyl substituted with at least onehydroxyl group, such as —CH₂OH, —CH(OH)CH₃, and the like.

“Mono- or di(C₃₋₆cycloalkyl)methyl” represents a methyl groupsubstituted with one or two C₃₋₆cycloalkyl groups, such ascyclopropylmethyl, dicyclopropylmethyl, and the like.

“C₁₋₆alkylcarbonylC₁₋₆alkyl” represents a C₁₋₆alkyl substituted with a—COC₁₋₆alkyl group.

“C₁₋₆alkylcarbonyloxyC₁₋₆alkyl” represents a C₁₋₆alkyl substituted witha —COOC₁₋₆alkyl group.

“C₁₋₆alkyloxyC₁₋₆alkyl” represents a C₁₋₆alkyl substituted with a—OC₁₋₆alkyl group.

“C₁₋₆alkylthioC₁₋₆alkyl” represents a C₁₋₆alkyl substituted with a—SC₁₋₆alkyl group.

“Sulfanyl(C₁₋₆alkyl)” means —SO₂(C₁₋₆alkyl), such as —SO₂ methyl and thelike.

“Mono- or di(C₁₋₆alkyl)amino represents an amino substituted with oneC₁₋₆alkyl or with two C₁₋₆alkyls, respectively.

“(C₁₋₆alkyl)(C₁₋₆alkanoyl)amino” represents an amino substituted with aC₁₋₆alkyl and a C₁₋₆alkanoyl (i.e., C(═O)(C₁₋₆alkyl).

“Mono- or di(C₁₋₆alkyl)aminoC₁₋₆alkyl” represents a C₁₋₆alkylsubstituted with a mono- or di(C₁₋₆alkyl)amino.

“C₁₋₆alkylidenyl” represents a divalent C₁₋₆alkyl radical, such asmethylene (—CH₂—), ethylene (—CH₂CH₂—), and the like.

“C₁₋₆alkylidenyl having one, two or three carbon atoms replaced with aheteroatom individually selected from oxygen, nitrogen or and sulfur”means a C₁₋₆alkylidenyl wherein one, two or three methylenyl groups(i.e., “CH₂”) is replaced with O, N or S, such as —OCH₂O—, —OCH₂CH₂O—,and the like.

“C₃₋₁₂heterocycle” represents a ring made up of more than one kind ofatom, and which contains 3 to 12 carbon atoms, such as pyridinyl,pyrimidinyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl,pyridazinyl, pyrazinyl, triazinyl (such as 1, 3, 5), pyrrolyl,thiopenyl, oxazolyl, isoxazoly, pyrrolinyl, pyrrolidinyl, piperidinyl,and the like, as well as heterocyclic rings fused to phenyl to form abicyclic ring, such as pyrolidinophenyl and the like.

“Halo” means fluoro, chloro, bromo or iodo.

As used in the context of this invention,

represents —CH₂CH₂ ⁻ or —CH═CH— optionally substituted with 1 or 2 Rsubstituents (i.e., when n=1 and m=0, 1 or 2), or —CH₂CH₂CH₂— optionallysubstituted with 1, 2 or 3 R substituents (i.e., when n=2 and m=0, 1, 2or 3). Accordingly, representative compounds of this invention include(but are not limited to) compounds having the following structures(I-1), (I-2), (I-3), (I-4), (I-5) and (I-6):

More specifically, and depending upon the choice of the X moiety,representative CRF receptor antagonists of this invention includecompounds having the following structures (Ia) and (Ib), respectively:

In one preferred embodiment, the CRF receptor antagonists of thisinvention have structure (Ia). In another preferred embodiment, the CRFreceptor antagonists of this invention have structure (Ib), wherein R′is hydrogen. Such compounds are represented by the following structures(I-1a), (I-1b), (I-4a) and (I-4b):

As noted above, R₁ is —C(H)_(0,1)(R₃)(R₄) which represents —CH(R₃)(R₄)and —C(R₃)(R₄). Representative embodiments in this regard include thefollowing R₁ moieties:

Similarly, when R₃ is keto, representative R₁ moieties include thefollowing:

Representative R₁ moieties in this regard include —C(═O)R₄, —C(═O)OR₄,—C(═O)NH₂, —C(═O)NH(C₁₋₆alkyl) and —C(═O)N(C₁₋₆alkyl)(C₁₋₆alkyl).

In the embodiment where the R₃ and R₄ groups of R₁ taken together form aC₃₋₈cycloalkyl, the resulting R₁ group has the structure:

Representative C₃₋₈cycloalkyls include cyclopropyl, cyclopentyl andcyclohexyl. Furthermore, when the C₃₋₈cycloalkyl is a C₅₋₇cycloalkyl,optionally substituted with one or more C₁₋₆alkyl groups, arepresentative R₁ moiety has the following structure:

wherein R₅ and R₆ are the same or different and independently selectedfrom a C₁₋₆alkyl, such as methyl or ethyl.

Similarly, in the embodiment where the R₃ and R₄ groups of R₁ takentogether form a C₅₋₈cycloalkyl fused to Ar, the resulting R₁ group hasthe structure:

including optionally substituted analogs thereof as defined above.

In more specific embodiments of this invention, representative Ar groupsof this invention include 2,4,6-trimethylphenyl,2-chloro-4-methylphenyl, 2-chloro-4-methoxyphenyl,2-bromo-4-methylphenyl, 2-methyl-4-chlorophenyl, 2-methyl-4-bromophenyl,2-bromo-4-isopropylphenyl, 2,4-dichlorophenyl,2,6-dimethyl-4-bromophenyl, 4-chlorophenyl, 2,4-dimethoxyphenyl,2,4-dimethylphenyl, 2-methoxyphenyl, 3-methoxyphenyl,2-methyl-4-methoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyhl,4-trifluoromethylphenyl, 4-methoxyphenyl, 2,4,6-trifluorophenyl,2-methyl-4-N(ethyl)₂phenyl, 2-bromo-4-(OCF₃)phenyl,4-dimethylamino-2-methyl-3-pyridinyl,4-dimethylamino-6-methyl-2-pyridinyl, 4-dimethylamino-3-pyridinyl.4-N(CH₃)(COCH₃)-phenyl, 3,4-methylenedioxyphenyl and3,4-ethylenedioxyphenyl.

Representative optional R groups of this invention include methyl,ethyl, n-propyl, iso-propyl, iso-butyl, ═CH₂ and ═CHCH₃.

Representative R′ groups are hydrogen, fluoro, chloro, bromo, methyl andethyl, and preferably hydrogen.

Representative R₁ groups include methyl, ethyl, n-propyl, iso-propyl,n-butyl, iso-butyl, tert-butyl, n-pentyl, iso-pentyl, neo-pentyl,—CH(ethyl)₂, —CH(n-propyl)₂, —CH(n-butyl)₂, —CH₂CH₂OCH₃,—CH(methyl)(CH₂OCH₃), —CH(ethyl)(CH₂OCH₃), —CH(n-propyl)(CH₂OCH₃),—CH(n-butyl)(CH₂OCH₃), —CH(tert-butyl)(CH₂OCH₃), —CH(CH₂OCH₃)₂,—CH(benzyl)(CH₂OCH₃), —CH(4-chlorobenzyl)(CH₂OCH₃),—CH(CH₂OCH₃)(CH₂CH₂SCH₃), —CH(ethyl)(CH₂Obenzyl), —CHC≡CH,—CH(methyl)(ethyl), —CH(methyl)(n-propyl), —CH(methyl)(n-butyl),—CH(methyl)(n-pentyl), —CH(methyl)(CH₂CH₂CH₂CH(CH₃)₂),—CH(ethyl)(n-propyl), —CH(ethyl)(n-butyl), —CH(ethyl)(n-pentyl), ),—CH(n-propyl)(n-butyl), —CH(n-propyl)(n-pentyl), cyclopropyl,cyclobutyl, cyclohexyl, 2-methylcyclohexyl, 3-methylcyclohexyl,1,2,3,4-tetrahydronaphthyl (1 and 2), benzyl, 2-chlorobenzyl,—CH(methyl)(benzyl), —CH(ethyl)(benzyl), —CH(n-propyl)(benzyl),—CH(n-butyl)(benzyl), —CH₂(cyclopropyl), —CH₂(cyclobutyl),—CH₂CH(methyl)CH₂CH₃, —CH₂CH(ethyl)CH₂CH₃, —CH₂C(methyl)₃, —CH₂C≡CH,—CH₂C(═O)ethyl, —C(═O)cyclopropyl, —C(═O)NHbenzyl, —C(═O)methyl,—C(═O)benzyl, —C(═O)phenyl, —C(═O)ethyl, —C(═O)CH₂C(═O)Oethyl,—C(═O)CH(phenyl)ethyl, C(═O)pyridyl, —C(═O)(4-N,N-dimethylamino)phenyl,—C(═O)CH₂Omethyl, —C(═O)CH(ethyl)₂, —C(═O)n-butyl,—C(═O)CH₂CH₂(methyl)₂, —C(═O)n-propyl, —C(═O)CH₂CH₂phenyl, —CH₂pyridyl,—CH₂CH₂NHphenyl, —CH₂CH₂C(═O)Oethyl, —CH₂CH₂CH₂phenyl,—CH₂CH₂-N-phthalimide, —CH₂CH₂CH₂C(═O)Oethyl, —CH₂CH₂Oethyl,—CH₂CH(methyl)₂, —CH₂C(═O)Oethyl, —CH₂C(═O)pyrrohdinophenyl,—CH₂CH₂Ophenyl, —CH₂CH₂CH₂CH₂-N-phthalimide, —CH₂C(═O)Ot-butyl,—CH₂CH₂CH(methyl)₂, —CH₂C(═O)NH₂, —CH₂-4-(SO₂CH₃)phenyl, —CH₂CH₂pyrolyland benzyl.

Representative R₂ groups include methyl, ethyl and hydrogen, andpreferably methyl.

The compounds of the present invention may be prepared by known organicsynthesis techniques, including the methods described in more detail inthe Examples, and may generally be utilized as the free base.Alternatively, the compounds of this invention may be used in the formof acid addition salts. Acid addition salts of the free base aminocompounds of the present invention may be prepared by methods well knownin the art, and may be formed from organic and inorganic acids. Suitableorganic acids include maleic, fumaric, benzoic, ascorbic, succinic,methanesulfonic, acetic, oxalic, propionic, tartaric, salicylic, citric,gluconic, lactic, mandelic, cinnamic, aspartic, stearic, palmitic,glycolic, glutamic, and benzenesulfonic acids. Suitable inorganic acidsinclude hydrochloric, hydrobromic, sulfuric, phosphoric, and nitricacids.

More specifically, the compounds of the structure (I) may be madeaccording to the procedures set forth in Examples 1 and 2, as well as bythe following general Reaction Scheme:

The effectiveness of a compound as a CRF receptor antagonist may bedetermined by various assay methods. Suitable CRF antagonists of thisinvention are capable of inhibiting the specific binding of CRF to itsreceptor and antagonizing activities associated with CRF. A compound ofstructure (I) may be assessed for activity as a CRF antagonist by one ormore generally accepted assays for this purpose, including (but notlimited to) the assays disclosed by DeSouza et al. (J. Neuroscience7:88, 1987) and Battaglia et al. (Synapse 1:572, 1987). As mentionedabove, suitable CRF antagonists include compounds which demonstrate CRFreceptor affinity. CRF receptor affinity may be determined by bindingstudies that measure the ability of a compound to inhibit the binding ofa radiolabeled CRF (e.g., [¹²⁵I]tyrosine-CFR) to its receptor (e.g.,receptors prepared from rat cerebral cortex membranes). The radioligandbinding assay described by DeSouza et al. (supra, 1987) provides anassay for determining a compound's affinity for the CRF receptor. Suchactivity is typically calculated from the IC₅₀ as the concentration of acompound necessary to displace 50% of the radiolabeled ligand from thereceptor, and is reported as a “K_(i)” value calculated by the followingequation: $K_{i} = \frac{{IC}_{50}}{1 + {L/K_{D}}}$

where L=radioligand and K_(D)=affinity of radioligand for receptor(Cheng and Prusoff, Biochem. Pharmacol. 22:3099, 1973).

In addition to inhibiting CRF receptor binding, a compound's CRFreceptor antagonist activity may be established by the ability of thecompound to antagonize an activity associated with CRF. For example, CRFis known to stimulate various biochemical processes, including adenylatecyclase activity. Therefore, compounds may be evaluated as CRFantagonists by their ability to antagonize CRF-stimulated adenylatecyclase activity by, for example, measuring cAMP levels. TheCRF-stimulated adenylate cyclase activity assay described by Battagliaet al. (supra, 1987) provides an assay for determining a compound'sability to antagonize CRF activity. Accordingly, CRF receptor antagonistactivity may be determined by assay techniques which generally includean initial binding assay (such as disclosed by DeSouza (supra, 1987))followed by a cAMP screening protocol (such as disclosed by Battaglia(supra, 1987)).

With reference to CRF receptor binding affinities, CRF receptorantagonists of this invention have a K_(i) of less than 10 μM. In apreferred embodiment of this invention, a CRF receptor antagonist has aK_(i) of less than 1 μM, and more preferably less than 0.25 μM (i.e.,250 nM). As set forth in greater detail below, representative compoundsof this invention were assayed by the method of Example 4. Preferredcompounds having a K_(i) of less than 1 μM are compounds numbers (I-1)through (I-25) and (I-29) through (I-33). More preferred compoundshaving a K_(i) of less than 250 nM are compound numbers (I-1) through(I-14), (I-16) through (I-25) and (I-29) through (I-32).

The CRF receptor antagonists of the present invention demonstrateactivity at the CRF receptor site, and may be used as therapeutic agentsfor the treatment of a wide range of disorders or illnesses includingendocrine, psychiatric, and neurologic disorders or illnesses. Morespecifically, the CRF receptor antagonists of the present invention maybe useful in treating physiological conditions or disorders arising fromthe hypersecretion of CRF. Because CRF is believed to be a pivotalneurotransmitter that activates and coordinates the endocrine,behavioral and automatic responses to stress, the CRF receptorantagonists of the present invention can be used to treatneuropsychiatric disorders. Neuropsychiatric disorders which may betreatable by the CRF receptor antagonists of this invention includeaffective disorders such as depression; anxiety-related disorders suchas generalized anxiety disorder, panic disorder, obsessive-compulsivedisorder, abnormal aggression, cardiovascular abnormalities such asunstable angina and reactive hypertension; and feeding disorders such asanorexia nervosa, bulimia, and irritable bowel syndrome. CRF antagonistsmay also be useful in treating stress-induced immune suppressionassociated with various diseases states, as well as stroke. Other usesof the CRF antagonists of this invention include treatment ofinflammatory conditions (such as rheumatoid arthritis, uveitis, asthma,inflammatory bowel disease and G.I. motility), Cushing's disease,infantile spasms, epilepsy and other seizures in both infants andadults, and various substance abuse and withdrawal (includingalcoholism).

In another embodiment of the invention, pharmaceutical compositionscontaining one or more CRF receptor antagonists are disclosed. For thepurposes of administration, the compounds of the present invention maybe formulated as pharmaceutical compositions. Pharmaceuticalcompositions of the present invention comprise a CRF receptor antagonistof the present invention (i.e., a compound of structure (I)) and apharmaceutically acceptable carrier and/or diluent. The CRF receptorantagonist is present in the composition in an amount which is effectiveto treat a particular disorder—that is, in an amount sufficient toachieve CRF receptor antagonist activity, and preferably with acceptabletoxicity to the patient. Preferably, the pharmaceutical compositions ofthe present invention may include a CRF receptor antagonist in an amountfrom 0.1 mg to 250 mg per dosage depending upon the route ofadministration, and more preferably from 1 mg to 60 mg. Appropriateconcentrations and dosages can be readily determined by one skilled inthe art.

Pharmaceutically acceptable carrier and/or diluents are familiar tothose skilled in the art. For compositions formulated as liquidsolutions, acceptable carriers and/or diluents include saline andsterile water, and may optionally include antioxidants, buffers,bacteriostats and other common additives. The compositions can also beformulated as pills, capsules, granules, or tablets which contain, inaddition to a CRF receptor antagonist, diluents, dispersing and surfaceactive agents, binders, and lubricants. One skilled in this art mayfurther formulate the CRF receptor antagonist in an appropriate manner,and in accordance with accepted practices, such as those disclosed inRemington's Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co.,Easton, Pa. 1990.

In another embodiment, the present invention provides a method fortreating a variety of disorders or illnesses, including endocrine,psychiatric and neurologic disorders or illnesses. Such methods includeadministering of a compound of the present invention to a warm-bloodedanimal in an amount sufficient to treat the disorder or illness. Suchmethods include systemic administration of a CRF receptor antagonist ofthis invention, preferably in the form of a pharmaceutical composition.As used herein, systemic administration includes oral and parenteralmethods of administration. For oral administration, suitablepharmaceutical compositions of CRF receptor antagonists include powders,granules, pills, tablets, and capsules as well as liquids, syrups,suspensions, and emulsions. These compositions may also includeflavorants, preservatives, suspending, thickening and emulsifyingagents, and other pharmaceutically acceptable additives. For parentaladministration, the compounds of the present invention can be preparedin aqueous injection solutions which may contain, in addition to the CRFreceptor antagonist, buffers, antioxidants, bacteriostats, and otheradditives commonly employed in such solutions.

As mentioned above, administration of a compound of the presentinvention can be used to treat a wide variety of disorders or illnesses.In particular, the compounds of the present invention may beadministered to a warm-blooded animal for the treatment of depression,anxiety disorder, panic disorder, obsessive-compulsive disorder,abnormal aggression, unstable angina, reactive hypertension, anorexianervosa, bulimia, irritable bowel syndrome, stress-induced immunesuppression, stroke, inflammation, Cushing's disease, infantile spasms,epilepsy, and substance abuse or withdrawal.

The following examples are provided for purposes of illustration, notlimitation.

EXAMPLES

The CRF receptor antagonists of this invention may be prepared by themethods disclosed in Examples 1-2. Example 3 discloses representativecompounds of this invention. Example 4 presents a method for determiningthe receptor binding activity (K_(i)), and Example 5 discloses an assayfor screening compounds of this invention for CRF-stimulated adenylatecyclase activity.

Example 1 Synthesis of Representative Compounds of Structure (Ia)

Compound(4)

A solution of 4,6-dichloro-2-methyl-5-nitropyrimidine (3; J. Chem. Soc.1954, 3836) (2.23 g, 11 mmol) in EtOH (30 mL) at −30° C. was treatedwith 1-ethylpropylamine (870 mg, 10 mmol) in EtOH (8 mL) and thereaction mixture was stirred at −30° C. for 1 hour and then warmed toambient temperature. Volatiles were evaporated and the residue waspartitioned between water and EtOAc. The organic layer was dried (sodiumsulfate), evaporated, purified by flash chromatography (silica) to givecompound (4).

Compound (5)

A solution of compound (4) (2.07 g, 8 mmol) in acetonitrile (15 mL) wastreated with 2,4,6-trimethylaniline (1.35 g, 10 mmol) at ambienttemperature, then triethylamine (1.52 g, 15 mmol) was introduced. Thereaction mixture was stirred at ambient temperature for 2 hours.Volatiles were evaporated and the residue was partitioned between brineand EtOAc. The organic layer was dried (sodium sulfate), evaporated,purified by flash chromatography (silica) to give compound (5).

Compound (6)

Compound (5) (2.14 g, 6 mmol) was dissolved in 1:1 dioxane/water (20mL), and treated with concentrated aqueous ammonia hydroxide (5 mL).Sodium hydrosulfite (3.12 g, 18 mmol) was added in small batches overone hour and the solution was stirred at ambient temperature for 8hours. The reaction mixture was partitioned between brine and EtOAc. Theorganic layer was dried (sodium sulfate), evaporated, purified by flashchromatography (silica) to give compound (6).

Compound (7)

A mixture of compound (6) (654 mg, 2 mmol) and triethylamine (500 mg) indry THF (10 mL) was treated with triphosgene (217 mg, 0.73 mmol), andthe reaction mixture was stirred at ambient temperature for 1 hour.Precipitates were filtered and the filtrate was evaporated, and theresidue was partitioned between brine and EtOAc. The organic layer wasdried (sodium sulfate), evaporated, purified by flash chromatography(silica) to give compound (7).

Compound (8)

Compound (7) (353 mg, 1 mmol) in dry DMF (5 mL) was treated with NaH(120 mg, 3 mmol, 60% in oil) at ambient temperature. Then1,2-dibromoethane (654 mg, 3 mmol) was added to the reaction mixture andstirred for 10 hours. The reaction mixture was partitioned between waterand EtOAc. The organic layer was dried (sodium sulfate), evaporated,purified by flash chromatography (silica) to give compound (8). LC-MS380 (MH+).

Compound (9)

A solution of compound (8) (38 mg, 0.1 mmol) in toluene (2 ml) wastreated with activated manganese dioxide catalyst (100 mg) at reflux for16 hours. The catalyst was removed by filtration through a Celite padand the filtrate was evaporated to dryness and purified by Prepative TLC(silica gel) with ethyl acetate hexane (1:1) to provide compound (9).

Example 2 Synthesis of Representative Compounds of Structure (IB)

Compounds of structure (Ib) may be made by the same synthetic route asdisclosed above in Example 1, but employing the corresponding pyridineto compound (1) rather than the pyrimidine. For example, representativecompounds of this invention may be made by the following reactionscheme:

Example 3 Synthesis of Representative Compounds

Further representative compounds of this invention were made by generalReaction Scheme disclosed above and/or by the procedures of Examples 1and 2, and are presented in the following Table.

TABLE Representative Compounds

Cpd R X R₁ Ar (I-1) H N —CH(CH₂CH₂CH₃)₂ 2,4,6-trimethylphenyl (I-2) H CH—CH(CH₂CH₂CH₃)₂ 2-chloro-4-methylphenyl (I-3) H CH —CH(CH₂CH₂CH₃)₂2-bromo-4-isopropylphenyl (I-4) H CH —CH(CH₂CH₂CH₃)₂ 2,4-dichlorophenyl(I-5) H CH —CH(CH₂CH₂CH₃)₂ 2,6-dimethyl-4-bromophenyl (I-6) H CH—CH(CH₂CH₂CH₃)₂ 2-methyl-4-chlorophenyl (I-7) H CH —CH(CH₂CH₂CH₃)₂2-bromo-4-methylphenyl (I-8) H CH —CH(CH₂CH₂CH₃)₂ 4-chlorophenyl (I-9) HCH —CH(CH₂CH₂CH₃)₂ 2,4-dimethoxyphenyl (I-10) H CH —CH(CH₂CH₂CH₃)₂2-methoxyphenyl (I-11) H CH —CH(CH₂CH₂CH₃)₂ 3,4-dimethoxyphenyl (I-12) HCH —CH(CH₂CH₂CH₃)₂ 4-trifluoromethylphenyl (I-13) H CH —CH(CH₂CH₂CH₃)₂4-methoxyphenyl (I-14) H CH —CH(CH₂CH₂CH₃)₂ 2,4,6-trifluorophenyl (I-15)H CH —CH(CH₂CH₂CH₃)₂ 2-methyl-4- (diethylamine)phenyl (I-16) H CH—CH(CH₂CH₂CH₃)₂

(I-17) H CH —CH(CH₂CH₂CH₃)₂ 2-bromo-4-(OCF₃)phenyl (I-18) H CH—CH(CH₂CH₂CH₃)₂ 3-methoxyphenyl (I-19) H CH —CH(CH₂CH₂CH₃)₂2,4-dimethylphenyl (I-20) H CH —CH(CH₂CH₂CH₃)₂

(I-21) H CH —CH(CH₂CH₃)((CH₂)₃CH₃) 2-methyl-4-chlorophenyl (I-22) H CH—CH(CH₂CH₃)₂ 2-methyl-4-chlorophenyl (I-23) H CH —CH(CH₂CH₃)((CH₂)₂CH₃)2-methyl-4-chlorophenyl (I-24) H CH —CH(CH₂CH₃)((CH₂)₄CH₃)2-methyl-4-chlorophenyl (I-25) H CH —CH((CH₂)₂CH₃)((CH₂)₃CH₃)2-methyl-4-chlorophenyl (I-26) H CH —CH(CH₂CH₂CH₃)₂ 3,5-dimethoxyphenyl(I-27) H CH —CH(CH₂CH₂CH₃)₂ 3-(5-methylisoxazolyl) (I-28) H CH—CH(CH₂CH₂CH₃)₂ 4-phenoxyphenyl (I-29) H CH —CH(CH₂CH₂CH₃)₂4-methoxy-3-pyridinyl (I-30) H CH —CH(CH₂CH₂CH₃)₂4-dimethylamine-3-pyridinyl (I-31) ethyl(S) CH —CH₂CH₂OCH₃4-methoxyphenyl (I-32) ethyl(S) CH —CH₂CH₂OH 4-methoxyphenyl (I-33) H CH—CH(CH₂CH₂CH₃)₂ 4-(N-methyl-N-acetyl)phenyl (I-34) ethyl(S) CH—CH₂CH₂N(CH₂CH₃)₂ 4-methoxyphenyl (I-35) H CH —CH(CH₂CH₂CH₃)₂4-(COOMe)phenyl (I-36) H CH —CH(CH₂CH₂CH₃)₂ 4-chloro-3-pyridinyl (I-37)H CH —CH(CH₂CH₂CH₃)₂ 4-(COMe)phenyl (I-38) H CH —CH(CH₂CH₂CH₃)₂4-(CH(OH)(CH₃)₂)phenyl (I-39) ethyl(S) CH C(O)CH₃ 4-methoxyphenyl (I-40)ethyl(S) CH C(O)CH₂Ph 4-methoxyphenyl (I-41) ethyl(S) CH C(O)Ph4-methoxyphenyl (I-42) ethyl(S) CH C(O)CH₂CH₃ 4-methoxyphenyl (I-43)ethyl(S) CH C(O)CH₂CO₂CH₂CH₃ 4-methoxyphenyl (I-44) ethyl(S) CHC(O)Cyclopropyl 4-methoxyphenyl (I-45) ethyl(S) CH C(O)CH(Ph)CH₂CH₃4-methoxyphenyl (I-46) ethyl(S) CH C(O)4-Pyridyl 4-methoxyphenyl (I-47)ethyl(S) CH C(O)4-(N,N- 4-methoxyphenyl Dimethylamino)phenyl (I-48)ethyl(S) CH C(O)CH₂OCH₃ 4-methoxyphenyl (I-49) ethyl(S) CH C(O)3-Pyridyl4-methoxyphenyl (I-50) ethyl(S) CH C(O)CH(CH₂CH₃)CH₂CH₃ 4-methoxyphenyl(I-51) ethyl(S) CH C(O)CH₂CH₂CH₂CH₃ 4-methoxyphenyl (I-52) ethyl(S) CHC(O)CH₂CH(CH₃)₂ 4-methoxyphenyl (I-53) ethyl(S) CH C(O)CH₂CH₂CH₃4-methoxyphenyl (I-54) ethyl(S) CH C(O)CH₂CH₂Ph 4-methoxyphenyl (I-55)ethyl(S) CH CH₂-3-Pyridyl 4-methoxyphenyl (I-56) ethyl(S) CH CH₂CH₂NHPh4-methoxyphenyl (I-57) ethyl(S) CH CH₂CH₂CO₂CH₂CH₃ 4-methoxyphenyl(I-58) ethyl(S) CH CH₂CH₂CH₂Ph 4-methoxyphenyl (I-59) ethyl(S) CHCH₂CH₂—N-Phthalimide 4-methoxyphenyl (I-60) ethyl(S) CHCH₂CH₂CH₂CO₂CH₂CH₃ 4-methoxyphenyl (I-61) ethyl(S) CH CH₂CH₂OCH₂CH₃4-methoxyphenyl (I-62) ethyl(S) CH CH₂CH(CH₃)₂ 4-methoxyphenyl (I-63)ethyl(S) CH CH₂CO₂CH₂CH₃ 4-methoxyphenyl (I-64) ethyl(S) CHCH₂C(O)(4-Pyrrolidinophenyl) 4-methoxyphenyl (I-65) ethyl(S) CHCH₂CH₂OPh 4-methoxyphenyl (I-66) ethyl(S) CH CH₂CH₂CH₂CH₂—N-4-methoxyphenyl Phthalimide (I-67) ethyl(S) CH CH₂CO₂tBu 4-methoxyphenyl(I-68) ethyl(S) CH CH₂CH₂CH(CH₃)₂ 4-methoxyphenyl (I-69) ethyl(S) CHCH₂C(O)NH₂ 4-methoxyphenyl (I-70) ethyl(S) CH CH₂-4-(SO₂CH₃)Ph4-methoxyphenyl (I-71) ethyl(S) CH CH₂CH₂-1-Pyrrole 4-methoxyphenyl(I-72) ethyl(S) CH CH₂Ph 4-methoxyphenyl Cpd Analytical data (MS/¹H NMR)(I-1) 0.92 (t, 6H), 1.25-1.34 (m, 4H), 1.53-1.72 (m, 4H), 2.11 (s, 6H),2.31 (s, 3H), 2.46 (s, 3H), 3.51 (t, 2H), 4.02 (t, 2H), 4.37-4.42 (m,1H), 6.98 (s, 2H) (I-2) 0.93 (t, 6H), 1.21-1.38 (m, 4H), 1.52-1.60 (m,2H), 1.64-1.75 (m, 2H), 2.34 (s, 3H), 2.49 (s, 3H), 3.41 (t, 2H),3.72-3.79 (m, 1H), 3.92-4.12 (m, 2H), 6.25 (s, 1H), 7.05-7.78 (m, 3H)(I-3) 0.93 (t, 6H), 1.26 (d, 6H), 1.27-1.35 (m, 4H), 1.52-1.61 (m, 4H),2.41 (s, 3H), 2.94 (hept, 1H), 3.40 (t, 2H), 3.73-3.82 (m, 1H),3.89-4.11(m, 2H), 6.26 (s, 1H), 7.27-7.58 (m, 3H) (I-4) 0.93 (t, 6H),1.25-1.38 (m, 4H), 1.52-1.62 (m, 4H), 2.40 (s, 3H), 3.40 (t, 2H),3.72-3.78 (m, 1H), 3.90-4.11 (m, 2H), 6.26 (s, 1H), 7.26-7.57 (m, 3H)(I-5) LC/MS 471 (M + H) (I-6) 0.93 (t, 6H), 1.25-1.39 (m, 4H), 1.51-1.62(m, 4H), 2.44 (s, 3H), 3.38 (t, 2H), 3.72-3.79 (m, 1H), 3.83 (s, 3H),3.99 (t, 2H), 6.25 (s, 1H), 6.99-7.61 (m, 4H); MS (CI) m/z 395.10 (MH⁺);HRMS (FAB) m/z 417.2277 (100; MNa⁺ [C₂₃H₃₀N₄O₂Na] = 417.2266). (I-7)7.50 (d, 1H), 7.38 (d, 1H), 7.16 (dd, 1H), 6.42 (s, 1H), 4.28-4.39 (m,1H), 4.09 (t, 2H), 3.81 (t, 2H), 1.92 (s, 6H), 1.88-2.01 (m, 2H),1.64-1.75 (m, 2H), 1.21-1.35 (m, 4H), 0.91 (t, 6H). (I-8) 7.72 (d, 2H),7.38 (d, 2H), 6.20 (s, 1H), 3.92 (t, 2H), 3.63-3.72 (m, 1H), 3.32 (t,2H), 2.38 (s, 3H), 1.40-1.57 (m, 4H), 1.17-1.32 (m, 4H), 0.86 (t, 6H).(I-9) LC/MS 425 (M + H) (I-10) LC/MS 395 (M + H) (I-11) LC/MS 425 (M +H) (I-12) LC/MS 433 (M + H) (I-13) LC/MS 395 (M + H) (I-14) 6.86 (d,1H), 6.83 (d, 1H), 6.26 (s, 1H), 4.01 (t, 2H)<3.69-3.80 (m, 1H), 3.40(t, 2H), 2.41 (s, 3H), 1.47-1.67 (m, 4H), 1.25-1.39 (m, 4H), 0.83 (t,6H). (I-15) 7.10(d, 1H), 6.53-6.56(m, 2H), 6.22(s, 1H), 3.95-4.10(m,2H), 3.70-3.82(m, 1H), 3.12-3.40(m, 6H), 2.40(s, 3H), 1.51-1.60(m, 4H),1.23-1.38(m, 4H), 1.16(t, 6H), 0.895-0.960(m, 6H) (I-16) 0.92 (t, 6H),1.28-1.41 (m, 4H), 1.46-1.59 (m, 4H), 2.43 (s, 3H), 3.37 (t, 2H),3.71-3.77 (m, 1H), 3.98 (t, 2H), 4.27 (s, 4H), 6.25 (s, 1H), 6.93-7.26(m, 3H); MS (CI) m/z 423.20 (MH⁺). (I-17) 7.61(d, 1H), 7.49 (d, 1H),7.31 (dd, 1H), 6.27 (s, 1H), 4.06-4.13 (dt, 1H), 3.89-3.97 (dt, 1H),3.71-3.78 (m, 1H), 3.42 (t, 2H), 2.41 (s, 3H), 1.53-1.63 (m, 4H),1.28-1.39 (m, 4H), 0.91-0.97 (m, 6H). (I-18) 7.39-7.45(m, 1H),6.10-7.076(m, 3H), 6.31(s, 1H), 4.04(t, 2H), 3.78-3.87(m, 4H), 3.52(t,2H), 2.54(s, 3H), 1.59-1.67(m, 4H), 1.21-1.42(m, 4H), 0.95(t, 6H) (I-19)7.09-7.21(m, 3H), 6.23(s, 1H), 3.93-4.15(m, 2H), 3.71-3.77(m, 1H),3.93(t, 2H), 2.39(s, 3H), 2.35(s, 3H), 1.49-1.65(m, 4H), 1.26-1.41(m,4H), 0.90- 0.96(m, 6H) (I-20) 6.99 (dd, 1H), 7.00 (d, 1H), 6.72 (d, 1H),6.07 (s, 1H), 5.82 (s, 2H), 3.81 (t, 2H), 3.51-3.61 (m, 1H), 3.20 (t,2H), 2.26 (s, 3H), 1.33-1.50 (m, 4H), 1.07- 1.21 (m, 4H), 0.75 (t, 6H).(I-21) 7.35(s, 1H), 7.277-7.281(m, 2H), 6.27(s, 1H), 3.95-4.07 (m, 2H),3.63(pentet, 1H), 3.41(t, 2H), 2.41(s, 3H), 2.25(s, 3H), 1.59-1.66(m,4H), 1.19-1.36(m, 4H), 0.87-0.97(m, 6H) (I-22) 7.35(s, 1H),7.277-7.281(m, 2H), 6.27(s, 1H), 3.93-4.06 (m, 2H), 3.51- 3.61(m, 1H),3.41(t, 2H), 2.41(s, 3H), 2.25(s, 3H), 1.59-1.70(m, 4H), 0.95(t, 3H),0.92(t, 3H) (I-23) 7.35(s, 1H), 7.275-7.28(m 2H), 6.26(s, 1H),3.91-4.1(m, 2H), 3.6-3.75(m, 1H), 3.41(t,, 2H), 2.41(s, 3H), 2.24(s,3H), 1.54-1.68(m, 4H), 1.30-1.42(m, 2H), 0.90-0.97(m, 6H) (I-24) 7.35(s,1H), 7.278-7.282(m, 2H), 6.26(s, 1H), 3.92-4.10(m, 2H), 3.624(pentet,1H), 3.41(t, 2H), 2.41(s, 3H), 2.25(s, 3H), 1.54-1.68(m, 4H),1.20-1.38(m, 6H), 0.84-0.97(m, 6H) (I-25) 7.35(s, 1H), 7.28(s, 2H),62.4(s, 1H), 3.92-4.10(m, 2H), 3.72(pentet, 1H), 3.40(t, 2H), 2.40(s,3H), 2.24(s, 3H), 1.56-1.62(m, 4H), 1.26-1.38(m, 6H), 0.86-0.97(m, 6H)(I-26) 7.01 (d, 2H), 6.42 (t, 1H), 6.26 (s, 1H), 4.00 (t, 2h), 3.80 (s,6H), 3.69-3.80 (m, 1H), 3.38 (t, 2H), 2.45 (s, 3H), 1.46-1.66 (m, 4H),1.23-1.38 (m, 4H), 0.92 (t, 6H). (I-27) 6.79 (d, 1H), 6.29 (s, 1H), 3.97(t, 2H), 3.69-3.79 (m, 1H), 3.37 (t, 2H), 2.51 (s, 3H), 2.47 (d, 3H),1.51-1.75 (m, 4H), 1.25-1.37 (m, 4H), 0.83-0.94 (m, 6H). (I-28) 7.72 (d,2H), 7.33-7.38 (m, 2H), 7.06-7.14 (m, 5H), 4.00 (t, 2H), 3.70-3.80 (m,1H), 3.39 (t, 2H), 2.45 (s, 3H), 1.47-1.65 (m, 4H), 1.25-1.39 (m, 4H),0.93 (t, 6H). (I-29) 8.57 (d, 1H), 7.95 (dd, 1H)<6.86 (d, 1H), 6.26 (s,1H), 4.00 (t, 2H), 3.91 (s, 3H), 3.70-3.79 (m, 1H), 3.39 (t, 2H), 2.43(s, 3H), 1.51-1.67 (m, 4H), 1.25- 1.38 (m, 4H), 0.92 (t, 6H). (I-30)0.92 (t, 6H), 1.25-1.38 (m, 4H), 1.51-1.62 (m, 4H), 2.42 (s, 3H); 3.12(s, 6H), 3.38 (t, 2H), 3.71-3.77 (m, 1H), 3.99 (t, 2H), 6.24 (s, 1H),6.63-8.47 (m, 3H); MS (CI) m/z 409.20 (MH⁺). (I-31) 7.66(d, 2H), 7.01(d,2H), 6.2(s, 1H), 4.18(dd, 1H), 3.84(s, 3H), 3.54-3.72(m, 5H),3.38-3.44(m, 1H), 3.37(s, 3H), 2.45(s, 3H), 1.31-1.73(m, 2H), 1.00(t,3H) (I-32) 7.62(d, 2H), 7.01(d, 2H), 6.23(s, 1H), 4.23(dd, 1H), 3.84(s,3H), 3.60-3.81(m, 5H), 3.31-3.40(m, 1H), 2.44(s, 3H), 1.5-1.72(m, 2H),1.01(t, 3H) (I-33) 7.94(d, 2H), 7.30(d, 2H), 6.29(s, 1H), 4.01(t, 2H),3.76(s, 1H), 3.41(t, 2H), 3.28(s, 3H), 2.48(s, 3H), 1.96(t, 3H),1.49-1.68(m, 4H), 1.25-1.37(m, 4H), 0.94(t, 6H) (I-34) 7.64(d, 2H),7.01(d, 2H), 6.2(s, 1H), 4.18(dd, 1H), 3.84(s, 3H), 3.56-3.74(m, 3H),3.22-3.32(m, 1H), 2.55-2.65(m, 6H), 2.45(s, 3H), 1.52-1.73(m, 2H),0.98-1.07(m, 9H) (I-35) δ 8.16 (d, 2H), 8.05 (d, 2H), 6.28 (s, 1H),4.00t, 2H), 3.93 (s, 3H), 3.71-3.81 (m, 1H), 3.39 (t, 2H), 2.47 (s, 3H),1.52-1.65 (m, 4H), 1.28-1.39 (m, 4H), 0.92 (t, 6H). (I-36) δ 8.60 (d,1H), 7.82 (dd, 1H), 7.74 (d, 1H), 6.29 (s, 1H), 3.99 (t, 2H), 3.73- 3.78(m, 1H), 3.87 (t, 2H), 2.48 (s, 3H), 1.51-1.62 (m, 4H), 1.25-1.38 (m,4H), 0.92 (t, 6H). (I-37) δ 8.08 (s, 4H), 6.29 (s, 1H), 4.00 (t, 2H),3.71-3.81 (m, 1H), 3.97 (t, 2H), 2.62 (s, 3H), 2.47 (s, 3H), 1.52-1.62(m, 4H), 1.22-1.39 (m, 4H), 0.93 (t, 6H). (I-38) δ 7.72 (d, 2H), 7.60(d, 2H), 6.27 (s, 1H), 4.00 (t, 2H), 3.68-3.82 (m, 1H), 3.48 (s, 6H),3.39 (t, 3H), 2.45 (s, 3H), 1.54-1.63 (m, 4H), 1.24-1.38 (m, 4H), 1.08(t, 3H), 0.93 (t, 3H).

Example 4 CRF Receptor Binding Activity

The compounds of this invention may be evaluated for binding activity tothe CRF receptor by a standard radioligand binding assay as generallydescribed by DeSouza et al. (J. Neurosci. 7:88-100, 1987). By utilizingvarious radiolabeled CRF ligands, the assay may be used to evaluate thebinding activity of the compounds of the present invention with any CRFreceptor subtype. Briefly, the binding assay involves the displacementof a radiolabeled CRF ligand from the CRF receptor.

More specifically, the binding assay is performed in 1.5 ml Eppendorftubes using approximately 1×10⁶ cells per tube stably transfected withhuman CRF receptors. Each tube receives about 0.1 ml of assay buffer(e.g., Dulbecco's phosphate buffered saline, 10 mM magnesium chloride,20 μM bacitracin) with or without unlabeled sauvagine, urotensin I orCRF (final concentration, 1 μM) to determine nonspecific binding, 0.1 mlof [¹²⁵I ] tyrosine-ovine CRF (final concentration ˜200 pM orapproximately the K_(D) as determined by Scatchard analysis) and 0.1 mlof a membrane suspension of cells containing the CRF receptor. Themixture is incubated for 2 hours at 22° C. followed by the separation ofthe bound and free radioligand by centrifugation. Following two washesof the pellets, the tubes are cut just above the pellet and monitored ina gamma counter for radioactivity at approximately 80% efficiency. Allradioligand binding data may be analyzed using the non-linearleast-square curve-fitting program LIGAND of Munson and Rodbard (Anal.Biochem. 107:220, 1990).

Example 5 CRF-Stimulated Adenylate Cyclase Activity

The compounds of the present invention may also be evaluated by variousfunctional testing. For example, the compounds of the present inventionmay be screened for CRF-stimulated adenylate cyclase activity. An assayfor the determination of CRF-stimulated adenylate cyclase activity maybe performed as generally described by Battaglia et al. (Synapse 1:572,1987), with modifications to adapt the assay to whole cell preparations.

More specifically, the standard assay mixture may contain the followingin a final volume of 0.5 ml: 2 mM L-glutamine, 20 mM HEPES, and 1 mMIMBX in DMEM buffer. In stimulation studies, whole cells with thetransfected CRF receptors are plated in 24-well plates and incubated for1 h at 37° C. with various concentrations of CRF-related and unrelatedpeptides in order to establish the pharmacological rank-order profile ofthe particular receptor subtype. Following the incubation, the media isaspirated, the wells rinsed once gently with fresh media, and the mediaaspirated. To determine the amount of intracellular cAMP, 300 μl of asolution of 95% ethanol and 20 mM aqueous hydrochloric acid is added toeach well and the resulting suspensions are incubated at −20° C. for 16to 18 hours. The solution is removed into 1.5 ml Eppendorf tubes and thewells washed with an additional 200 μl of ethanol/aqueous hydrochloricacid and pooled with the first fraction. The samples are lyophilized andthen resuspended with 500 μl sodium acetate buffer. The measurement ofcAMP in the samples is performed using a single antibody kit fromBiomedical Technologies Inc. (Stoughton, Mass.). For the functionalassessment of the compounds, a single concentration of CRF or relatedpeptides causing 80% stimulation of cAMP production is incubated alongwith various concentrations of competing compounds (10⁻¹² to 10⁻⁶ M).

It will be appreciated that, although specific embodiments of theinvention have been described herein for purposes of illustration,various modifications may be made without departing from the spirit andscope of the invention. Accordingly, the invention is not limited exceptas by the appended claims.

What is claimed is:
 1. A method for treating stroke in a warm-bloodedanimal in need thereof, comprising administering to the animal aneffective amount of a compound having the following structure:

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein:n is 1 m is 0, 1, 2 or 3; X is N or CR′; R′ is hydrogen, halogen or C₁₋₆alkyl; R is an optional substituent which, at each occurrence, isindependently C₁₋₆alkyl, C₃₋₆alkenyl, C₁₋₆alkylidenyl or C₁₋₆alkylAr; R₁is —C(H)_(0,1)(R₃)(R₄); R₂ is hydrogen or C₁₋₆alkyl; R₃ is hydrogen,keto, C₁₋₆alkyl, mono- or di(C₃₋₆cycloalkyl)methyl, C₁₋₆cycloalkyl,C₃₋₆alkenyl, hydroxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl, orC₁₋₆alkyloxyC₁₋₆alkyl, and R₄ is hydrogen, Ar¹, Ar¹C₁₋₆alkyl, OAr¹,C₁₋₈alkyl, C₁₋₆alkyloxy, C₃₋₆cycloalkyl, mono- ordi(C₃₋₆cycloalkyl)methyl, C₃₋₆alkenyl, C₃₋₆alkynyl,C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkoxyAr¹, hydroxyC₁₋₆alkyl,thienylC₁₋₆alkyl, furanylC₁₋₆alkyl, C₁₋₆alkylthioC₁₋₆alkyl, morpholinyl,mono- or di(C₁₋₆alkyl)aminoC₁₋₆alkyl, amino, (C₁₋₆alkyl)amino,di(C₁₋₆alkyl)amino, (C₁₋₆alkylAr¹)amino, (C₁₋₆alkyl)(Ar¹)amino,C₁₋₉alkylcarbonylC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,sulfonyl(C₁₋₈alkyl), C(═O)C₁₋₆alkyl, C₁₋₈alkyl substituted withphthalimide, Ar¹, OAr¹, NHAr¹, C(═O)Ar¹, C(═O)NHAr¹ or —C(═O)NH₂ or aradical of the formula —(C₁₋₆alkanediyl)-Y—(CO)_(0,1)—Ar¹ where Y is O,NH or a direct bond, or R₃ and R₄ taken together with the carbon atom towhich they are attached form a C₃₋₈cycloalkyl, a C₅₋₈cycloalkenyl, aC₃₋₁₂heterocycle, phenyl, naphthyl, or a C₅₋₈cycloalkyl fused to Ar¹,each of which being optionally substituted with one or more substituentsindependently selected from C₁₋₆alkyl; Ar is phenyl, naphthyl or anaromatic C₃₋₁₂heterocycle, each being optionally substituted with 1, 2or 3 substituents independently selected from halo, C₁₋₆alkyl,trifluoromethyl, O(trifluoromethyl), hydroxy, cyano, C₁₋₆alkyloxy,phenyoxy, benzoxy, C₁₋₆alkylthio, nitro, amino, mono- ordi(C₁₋₆alkyl)amino, (C₁₋₆alkyl)(C₁₋₆alkanoyl)amino, or piperidinyl, orwherein two substituents taken together are a C₁₋₆alkylidinyl or aC₁₋₆alkylidenyl having one, two or three carbon atoms replaced with aheteroatom individually selected from oxygen, nitrogen or sulfur; andAr¹ is phenyl, naphthyl or an aromatic C₃₋₁₂heterocycle, each of whichbeing optionally substituted with 1, 2 or 3 substituents independentlyselected from halo, C₁₋₆alkyl, C₁₋₆alkyloxy, di(C₁₋₆alkyl)amino,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, trifluoromethyl, sulfonyl(C₁₋₆alkyl), andC₁₋₆alkyl substituted with morpholinyl.
 2. A method for treatingdepression, anxiety disorder, panic disorder, obsessive-compulsivedisorder, abnormal aggression, unstable angina, reactive hypertension,anorexia nervosa, bulimia, irritable bowel syndrome, stress-inducedimmune suppression, inflammation, Cushing's disease, substance abuse orwithdrawal, infantile spasms, or epilepsy in a warm-blooded animal inneed thereof, comprising administering to the animal an effective amountof a compound having the following structure:

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein:n is 1; m is 0, 1, 2 or 3; X is N or CR′; R′ is hydrogen, halogen orC₁₋₆ alkyl; R is an optional substituent which, at each occurrence, isindependently C₁₋₆alkyl, C₃₋₆alkenyl, C₁₋₆alkylidenyl or C₁₋₆alkylAr; R₁is —C(H)_(0,1)(R₃)(R₄); R₂ is hydrogen or C₁₋₆alkyl; R₃ is hydrogen,keto, C₁₋₆alkyl, mono- or di(C₃₋₆cycloalkyl)methyl, C₃₋₆cycloalkyl,C₃₋₆alkenyl, hydroxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl, orC₁₋₆alkyloxyC₁₋₆alkyl, and R₄ is hydrogen, Ar¹, Ar¹C₁₋₆alkyl,OAr¹C₁₋₈alkyl, C₁₋₆alkyloxy, C₃₋₆cycloalkyl, mono- ordi(C₃₋₆cycloalkyl)methyl, C₃₋₆alkenyl, C₁₋₆alkynyl,C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkoxyAr¹, hydroxyC₁₋₆alkyl,thienylC₁₋₆alkyl, furanylC₁₋₆alkyl, C₁₋₆alkylthioC₁₋₆alkyl, morpholinyl,mono- or di(C₁₋₆alkyl)aminoC₁₋₆alkyl, amino, (C₁₋₆alkyl)amino,di(C₁₋₆alkyl)amino, (C₁₋₆alkylAr¹)amino, (C₁₋₆alkyl)(Ar¹)amino,C₁₋₆alkylcarbonylC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,sulfonyl(C₁₋₈alkyl), C(═O)C₁₋₆alkyl, C₁₋₈alkyl substituted withphthalimide, Ar¹, OAr¹, NHAr¹, C(═O)Ar¹, C(═O)NHAr¹ or —C(═O)NH₂, or aradical of the formula —(C₁₋₆alkanediyl)-Y—(CO)_(0,1)—Ar¹ where Y is O,NH or a direct bond, or R₃ and R₄ taken together with the carbon atom towhich they are attached form a C₃₋₈cycloalkyl, a C₅₋₈cycloalkenyl, aC₃₋₁₂heterocycle, phenyl, naphthyl, or a C₅₋₈cycloalkyl fused to Ar¹,each of which being optionally substituted with one or more substituentsindependently selected from C₁₋₆alkyl; Ar is phenyl, naphthyl or anaromatic C₃₋₁₂heterocycle, each being optionally substituted with 1, 2or 3 substituents independently selected from halo, C₁₋₆alkyl,trifluoromethyl, O(trifluoromethyl), hydroxy, cyano, C₁₋₆alkyloxy,phenyoxy, benzoxy, C₁₋₆alkylthio, nitro, amino, mono- ordi(C₁₋₆alkyl)amino, (C₁₋₆alkyl)(C₁₋₆alkanoyl)amino, or piperidinyl, orwherein two substituents taken together are a C₁₋₆alkylidinyl or aC₁₋₆alkylidenyl having one, two or three carbon atoms replaced with aheteroatom individually selected from oxygen, nitrogen or sulfur; andAr¹ is phenyl, naphthyl or an aromatic C₃₋₁₂heterocycle, each of whichbeing optionally substituted with 1, 2 or 3 substituents independentlyselected from halo, C₁₋₆alkyl, C₁₋₆alkyloxy, di(C₁₋₆alkyl)amino,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, trifluoromethyl, sulfonyl(C₁₋₆alkyl), andC₁₋₆alkyl substituted with morpholinyl.
 3. The method of any of claims 1or 2 wherein the compound has the structure


4. The method of any of claims 1 or 2 wherein the compound has thestructure


5. The method of any of claims 1 or 2 wherein m is
 0. 6. The method ofclaim 5 wherein the compound has the structure:


7. The method of claim 5 wherein the compound has the structure:


8. The method of any of claims 1 or 2 wherein m is
 1. 9. The method ofclaim 8 wherein the compound has the structure:


10. The method of claim 8 wherein the compound has the structure:


11. The method of any of claims 1 or 2 wherein X is CR′ and R′ ishydrogen.
 12. The method of any of claims 1 or 2 wherein X is N.
 13. Themethod of any of claims 1 or 2 wherein R is C₁₋₆alkyl.
 14. The method ofany of claims 1 or 2 wherein R is methyl or ethyl.
 15. The method of anyof claims 1 or 2 wherein R is ethyl.
 16. The method of any of claims 1or 2 wherein Ar is 2,4,6-trimethylphenyl, 2-chloro-4-methylphenyl,2-chloro-4-methoxyphenyl, 2-bromo-4-methylphenyl,2-methyl-4-chlorophenyl, 2-methyl-4-bromophenyl,2-bromo-4-isopropylphenyl, 2,4-dichlorophenyl,2,6-dimethyl-4-bromophenyl, 4-chlorophenyl, 2,4-dimethoxyphenyl,2,4-dimethylphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl,2-methyl-4-methoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl,4-trifluoromethylphenyl, 2,4,6-trifluorophenyl,2-methyl-4-N(ethyl)₂phenyl, 2-bromo-4-(OCF₃)phenyl,4-dimethylamino-2-methylpyridin-3-yl,4-dimethylamino-6-methylpyridin-3-yl, 4-dimethylamino-pyridin-3-yl,4-N(CH₃)(Ac)phenyl, 5-methylisoxazol-3-yl, 3,4-methylenedioxyphenyl, or3,4-ethylenedioxyphenyl.
 17. The method of any of claims 1 or 2 whereinAr is 2,4,6-trimethylphenyl, 2-methyl-4-chlorophenyl,2-chloro-4-methylphenyl, 2,4-dichlorophenyl, 2,6-dimethyl-4-bromophenyl,2-bromo-4-methylphenyl, 4-methoxyphenyl, or 4-chlorophenyl.
 18. Themethod of any of claims 1 or 2 wherein R₁ is methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, n-pentyl, iso-pentyl, neo-pentyl,—CH(ethyl)₂, —CH(n-propyl)₂, —CH(n-butyl)₂, —CH₂CH₂OCH₃,—CH(methyl)(CH₂OCH₃), —CH(ethyl)(CH₂OCH₃), —CH(n-propyl)(CH₂OCH₃),—CH(n-butyl)(CH₂OCH₃), —CH(tert-butyl)(CH₂OCH₃), —CH(CH₂OCH₃)₂,—CH(benzyl)(CH₂OCH₃), —CH(4-chlorobenzyl)(CH₂OCH₃),—CH(CH₂OCH₃)(CH₂CH₂SCH₃), —CH(ethyl)(CH₂Obenzyl), —CH(methyl)(ethyl),—CH(methyl)(n-propyl), —CH(methyl)(n-butyl), —CH(methyl)(n-pentyl),—CH(methyl)(CH₂CH₂CH₂CH(CH₃)₂), —CH(ethyl)(n-propyl),—CH(ethyl)(n-butyl), —CH(ethyl)(n-pentyl), —CH(n-propyl)(n-butyl),—CH(n-propyl)(n-pentyl), cyclopropyl, cyclobutyl, cyclohexyl,2-methylcyclohexyl, 3-methylcyclohexyl, 1,2,3,4-tetrahydronaphthyl (1and 2), benzyl, 2-chlorobenzyl, —CH(methyl)(benzyl), —CH(ethyl)(benzyl),—CH(n-propyl)(benzyl), —CH(n-butyl)(benzyl), —CH₂(cyclopropyl),—CH₂(cyclobutyl), —CH₂CH(methyl)CH₂CH₃, —CH₂CH(ethyl)CH₂CH₃,—CH₂C(methyl)₃, —CH₂C≡CH, —CH₂C(═O)ethyl, —C(═O)cyclopropyl,—C(═O)NHbenzyl, —C(═O)methyl, —C(═O)benzyl, —C(═O)phenyl, —C(═O)ethyl,—C(═O)CH₂C(═O)Oethyl, —C(═O)CH(phenyl)ethyl, C(═O)pyridyl,—C(═O)(4-N,N-dimethylamino)phenyl, —C(═O)CH₂Omethyl, —C(═O)CH(ethyl)₂,—C(═O)n-butyl, —C(═O)CH₂CH₂(methyl)₂, —C(═O)n-propyl,—C(═O)CH₂CH₂phenyl, —CH₂pyridyl, —CH₂CH₂NHphenyl, —CH₂CH₂C(═O)Oethyl,—CH₂CH₂CH₂phenyl, —CH₂CH₂—N-phthalimide, —CH₂CH₂CH₂C(═O)Oethyl,—CH₂CH₂Oethyl, —CH₂CH(methyl)₂, —CH₂C(═O)pyrrolidinophenyl,—CH₂CH₂Ophenyl, —CH₂CH₂CH₂CH₂—N-phthalimide, —CH₂CH₂CH₂CH(methyl)₂,—CH₂C(═O)NH₂, —CH₂-4-(SO₂CH₃)phenyl, or —CH₂CH₂pyrrolyl.
 19. The methodof any of claims 1 or 2 wherein R₁ is —CH(ethyl)₂, —CH(n-propyl)₂,—CH(ethyl)(n-butyl), or —CH(ethyl)(n-pentyl).
 20. The method of claim 2for treating depression.
 21. The method of claim 2 for treating anxietydisorder.
 22. The method of claim 2 for treating panic disorder.
 23. Themethod of claim 2 for treating obsessive-compulsive disorder.
 24. Themethod of claim 2 for treating abnormal aggression.
 25. The method ofclaim 2 for treating unstable angina.
 26. The method of claim 2 fortreating reactive hypertension.
 27. The method of claim 2 for treatinganorexia nervosa.
 28. The method of claim 2 for treating bulimia. 29.The method of claim 2 for treating irritable bowel syndrome.
 30. Themethod of claim 2 for treating stress-induced immune suppression. 31.The method of claim 2 for treating inflammation.
 32. The method of claim2 for treating Cushing's disease.
 33. The method of claim 2 for treatingsubstance abuse or withdrawal.
 34. The method of claim 2 for treatinginfantile spasms.
 35. The method of claim 2 for treating epilepsy.